GSM5750625: AID-TRRAP-cloneAT63-NaOH-12h-HA-CR21; Homo sapiens; OTHER
CUT&RUN experiments were implemented based on the procedure described by Meers et al, 2019 (doi: 10.7554/eLife.46314). Briefly, 250,000 cells were immobilized on 10ul of concavalin A-coated beads, permeabilized with digitonin, and incubated overnight at 4°C with an anti-HA tag antibody. The pA-MNase recombinant fusion protein was incubated at 4°C for 1h, at a final concentration of 700 ng/mL and then activated with CaCl2 during 30 min at 0°C. The reaction is stopped and DAN fragments are resealed by incubation at 37°C for 15min. Library were then constructed from released DNA fragments purified on NucleoSpin columns (Macherey-Nagel). Libraries for Illumina sequencing were prepared with KAPA Single-Indexed adapters set A&B and the Hyper Prep kit (KAPA Biosystems).